Electrostatic nanoparticles and use thereof

Abstract

The present invention relates to a method of generating a nanoparticle comprising (c) contacting an antibody with a composition comprising a first conjugate (A), the first conjugate comprising a positively charged polypeptide conjugated to a bifunctional linker, characterized in that the composition is essentially free of unconjugated bifunctional linker, thereby obtaining a second conjugate (B), the second conjugate comprising the positively charged polypeptide, the bifunctional linker, and the antibody; and (d) contacting the second conjugate (B), a positively charged polypeptide, and a negatively charged molecule, thereby forming a nanoparticle. The present invention also relates to a nanoparticle obtainable by a method of the invention, as well as to a nanoparticle comprising (a) a positively charged polypeptide; (b) a second conjugate (B), the second conjugate comprising an antibody conjugated to a positively charged polypeptide; (c) one or more negatively charged molecules. The present invention also relates to a composition comprising a nanoparticle of the invention and to a nanoparticle or composition of the invention for use in therapy.

1 . A method of generating a nanoparticle comprising
 c) contacting an antibody with a composition comprising a first conjugate (A), the first conjugate comprising a positively charged polypeptide conjugated to a bifunctional linker, characterized in that the composition is essentially free of unconjugated bifunctional linker, thereby obtaining a second conjugate (B), the second conjugate comprising the positively charged polypeptide, the bifunctional linker, and the antibody; and   d) contacting the second conjugate (B), a positively charged polypeptide, and a negatively charged molecule, thereby forming a nanoparticle.   
     
     
         2 . The method of  claim 1 , wherein the method comprises prior to step c):
 a) conjugating a positively charged polypeptide with a bifunctional linker;   b) removing unconjugated bifunctional linker.   
     
     
         3 . The method of  claim 1  or  2 , wherein the molar ratio between the first conjugate (A) and the antibody is at least about 10:1 in step c). 
     
     
         4 . The method of any one of the preceding claims, wherein the molar ratio between the positively charged polypeptide and the second conjugate (B) is at least about 10:1 in step d). 
     
     
         5 . The method of any one of the preceding claims, wherein the antibody is specific for a cell surface molecule. 
     
     
         6 . The method of any one of the preceding claims, wherein the negatively charged molecule is a nucleic acid. 
     
     
         7 . The method of any one of the preceding claims, wherein the positively charged polypeptide is a protamine or histone. 
     
     
         8 . A nanoparticle obtainable by a method of any one of the preceding claims. 
     
     
         9 . A nanoparticle comprising:
 a) a positively charged polypeptide;   b) a second conjugate (B), the second conjugate comprising an antibody conjugated to a positively charged polypeptide; and   c) one or more negatively charged molecule(s).   
     
     
         10 . The nanoparticle of  claim 8  or  9 , wherein the second conjugate is enriched in the outer portion of the nanoparticle. 
     
     
         11 . The nanoparticle of any one of  claims 8 - 10 , wherein the one or more negatively charged molecules are enriched in the inner portion of the nanoparticle. 
     
     
         12 . The nanoparticle of any one of  claims 8 - 11 , wherein the nanoparticle has a mean diameter of about 0.05 μm to about 10 μm. 
     
     
         13 . A composition comprising a nanoparticle of any one of  claims 8 - 12 . 
     
     
         14 . A nanoparticle of any one of  claims 8 - 12  or a composition of  claim 13  for use in therapy. 
     
     
         15 . A kit comprising a nanoparticle of any one of  claims 8 - 12  or a composition of  claim 13 .